Epigenome, MED12 and Progesterone Action in Uterine Leiomyomas

表观基因组、MED12 和孕酮在子宫平滑肌瘤中的作用

• 批准号: 10396486
• 负责人: Serdar E. Bulun
• 金额: $ 48.87万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10396486
• 关键词: Acetates; Affect; Age; Binding Sites; CRISPR/Cas technology; Cell Differentiation process; Cell Proliferation; Cells; ChIP-seq; Chromatin; Clonal Expansion; Collaborations; Common Neoplasm; Complement; Complex; Critical Pathways; DNA; Disease; Epigenetic Process; Etiology; Extracellular Matrix Proteins; Fibroid Tumor; Foundations; Gene Expression; Gene Mutation; Genes; Genetic Transcription; Genotype; Goals; Growth; Growth Factor; Health; Histones; Human; Knock-in Mouse; Knock-out; Leiomyoma; Maps; Mediating; Mediator of activation protein; Medical; Modification; Morbidity - disease rate; Mus; Mutate; Nucleic Acid Regulatory Sequences; Output; Paracrine Communication; Pathway interactions; Pharmacogenomics; Population; Progesterone; RNA; Regulatory Element; Research; Research Project Grants; Risk; Role; Signal Transduction; Site; Smooth Muscle Tumor; Supporting Cell; Symptoms; Testing; Tissues; Uterine Fibroids; Uterus; Woman; Work; antagonist; cytokine; driver mutation; effective therapy; epigenome; epigenomics; gain of function; gain of function mutation; genome-wide; histone modification; in vivo; liver injury; mouse model; myometrium; new therapeutic target; novel; paracrine; precision medicine; receptor; recruit; reproductive; response; stem; stem cell proliferation; stem cell survival; stem cells; transcriptome; transcriptome sequencing; tumor; tumor growth; tumorigenesis; tumorigenic

Uterine leiomyoma (LM, fibroid) is the most common tumor in women. No long-term medical treatment is available. Each LM seems to originate from the clonal expansion of a single mutated LM stem cell (LSC) in the myometrium (MYO). LSC comprise 5% of tumor mass and differentiate into an intermediate cell population (LIC, 7%), which then become terminally differentiated cells (LDC) comprising 88% of the tumor bulk. Driver mutations of mediator complex subunit 12 (mut-MED12) occur in 70% of all LM. Progesterone (P4) and its receptor PR are essential for LM growth. PR-rich LIC transduce P4 signaling to PR-deficient LSC via paracrine factors to activate their proliferation. Ulipristal acetate (UPA), a PR-selective P4 antagonist, shrank LM and reduced its symptoms, but its use was halted because of risk of liver injury. Our overall goal is to define the role of genomewide P4 action in the etiology of mut-MED12‒associated LM tumorigenesis and identify novel therapeutic targets. We found that mut-MED12 physically interacts with PR and genomewide PR-chromatin interaction landscapes are dramatically dysregulated in LM expressing mut-MED12 vs. normal MYO tissue carrying wild type MED12. Mut-MED12 enhances PR recruitment to cis-regulatory elements of P4 target genes encoding paracrine growth factors, cytokines and extracellular matrix proteins critical for LSC proliferation and LM tumorigenesis. Furthermore, the uteri of mice with a human-equivalent gain-of-function mutation in Med12 develop LM in response to P4, whereas Med12 knockout blocks P4/PR signaling in mouse uterus. We hypothesize that mut-MED12 alters PR-chromatin interaction signatures and enhances P4 action in LIC, providing a support niche for LSC survival and proliferation. Using ChIP-seq, RNA-seq, STARR-seq, and CRISPR/Cas9-gene editing strategies, and in vivo PDX and mut-Med12 knock-in mouse models, we propose the following Aims: (1) Determine whether PR-chromatin interaction loci specifically associated with mut- MED12 stimulate P4 action in a distinct stem-support cell population (LIC), which then send tumorigenic paracrine signals to increase LSC activity and tumor growth. We will test the hypothesis that mut-MED12 interacts with PR and alters its interaction with chromatin, thereby enhancing P4 responsiveness of LICs to activate gene transcription and paracrine signaling that support the function of adjacent LSC. (2) Define whether tumorigenic activity of mut-Med12 is mediated via altering PR-chromatin interaction landscapes in a human-equivalent mut-Med12 mouse model. We will test the hypothesis that mut-Med12 disrupts chromatin features surrounding PR-binding sites, thereby supporting gene transcription and pathways critical for LM tumorigenesis, whereas UPA shrinks LM via altering the aberrant PR-chromatin-epigenomic interactions and reversing inappropriate expression of disease-associated genes. Deciphering the genomewide mechanisms at a defined cell population level will help us identify genotype-specific novel targets associated with mut-MED12 for pharmacogenomics and precision medicine in the treatment of LM.

子宫平滑肌瘤（LM，肌瘤）是女性最常见的肿瘤。没有长期的医疗是 可用的。每个LM似乎源于单个突变的LM干细胞（LSC）的克隆膨胀 子宫肌（myo）。 LSC完成了5％的肿瘤质量，并分化为中间细胞种群 （LIC，7％），然后成为终止分化的细胞（LDC），完成了88％的肿瘤大块。司机 介体复合物亚基12（MUT-MED12）的突变发生在所有LM的70％中。孕酮（P4）及其 接收器PR对于LM生长至关重要。 Pr-Prich LIC通过旁分泌传递P4信号传导到PR缺陷LSC 激活其增殖的因素。乙酸乌统（UPA），PR选择P4拮抗剂LM和 降低了症状，但由于肝损伤的风险而停止使用。我们的总体目标是定义 全基因组P4作用在MUT-MED12 - 相关LM肿瘤发生的病因中的作用并鉴定出新的 治疗靶标。我们发现MUT-MED12与PR和全基因组PR染色质物理相互作用 相互作用景观在表达MUT-MED12与正常肌组织的LM中动态失调 携带野生型Med12。 MUT-MED12增强了PR募集到P4靶基因的顺式调节元件 编码旁分泌生长因子，细胞因子和细胞外基质蛋白对于LSC增殖至关重要 LM肿瘤发生。此外，在Med12中具有人类等效性突变的小鼠子宫 响应于P4的LM，而Med12敲除块中小鼠子宫中的P4/PR信号传导。我们 假设Mut-Med12改变了Pr-染色质的相互作用特征，并增强了LIC中的P4动作， 为LSC生存和增殖提供支持利基。使用chip-seq，RNA-Seq，Starr-Seq和 CRISPR/CAS9基因编辑策略以及体内PDX和MUT-MED12敲门鼠标模型，我们建议 以下目的：（1）确定pr染色质相互作用是否在局部特异性相关 MED12在独特的茎支持细胞种群（LIC）中刺激P4动作，然后发送小结核菌 旁分泌信号增加LSC活性和肿瘤生长。我们将测试mut-med12的假设 与PR相互作用并改变其与染色质的相互作用，从而增强了LIC对P4的反应性 激活支持相邻LSC功能的基因转录和旁分泌信号。 （2）定义 MUT-MED12的致瘤活性是否通过改变A中的Pr-染色质相互作用景观来介导 人类等效的MUT-MED12小鼠模型。我们将检验以下假设，即mut-med12破坏染色质 围绕PR结合位点的特征，从而支持LM至关重要的基因转录和途径 肿瘤发生，而UPA通过改变异常的染色质 - 基本相互作用和 逆转与疾病相关基因的不当表达。解密全基因组机制 定义的细胞种群水平将有助于我们确定与MUT-MED12相关的基因型特异性新靶 用于LM治疗的药物基因组学和精确医学。